Cerebral ischemia affects millions of Americans every year. Current treatments, although helpful, fail to prevent cognitive, motor, and speech impairment due to brain damage caused by cardiac arrest. The main goal of our proposal is the elucidation of the mechanisms by which ischemic preconditioning promotes neuroprotection against cerebral ischemia. We are developing cell-based therapies for cerebral vascular diseases offering a new paradigm for advancing and transforming the care of patients suffering global cerebral ischemia. The long-term goal of our labs is to prevent neurological consequences of cardiac arrest. In the current proposal we will focus on preventing hippocampal CA1 neuronal death due to cardiac arrest using principles developed from ischemic preconditioning in administering a subpopulation of highly homogeneous mesenchymal stromal cells (MSCs). We will assess the effectiveness of these cells utilizing well established brain slice cultures and animal model systems in our lab. Although MSCs are highly heterogeneous, we have developed the means of isolating a highly homogeneous subpopulation of MSCs. These cells were named marrow-isolated adult multilineage inducible (MIAMI) cells, based on their extensive proliferative capacity while maintaining telomere length and potential to generate mature cells derived from all three embryonic germ layers. Our previous studies demonstrate that human MIAMI cells differentiate to neuronal cells in vitro, to vascular endothelial cells in vitro and in vivo, and prevent tissue necrosis and reestablish blood flow in an animal model of peripheral vascular ischemia. To achieve our immediate goals, we will perform comparative effective research by comparing the effects of different stromal stem cell populations, cell conveying and engrafting approaches using novel technologies, comparing resveratrol-induced preconditioning with no preconditioning, and timing of cell delivery. However, any uses of cells for cellular therapy must address issues of cell delivery and engraftment. The use of pharmacologically active microcarriers (PAMs) conveying MIAMI cells could provide a powerful approach to resolve these issues and further repair lesioned tissues. PAMs are biodegradable, biocompatible poly(lactic-co-glycolic acid) microspheres that release therapeutic molecules in a controlled manner while providing a biomimetic surface and a 3D support in order to stimulate cell survival, orient and maintain their differentiation after transplantation. The scientific and technical challenge of the present project is to use resveratrol released by PAMs in a controlled manner to facilitate MIAMI cells and endogenous NSC survival and differentiation into neurons and to enhance endogenous neuroprotective, neurogenic, angiogenic and functional recovery mechanisms after cardiac arrest. We will pursue the following specific aims: Aim 1: Comparative effects of cell type and preconditioning a. Cell type: MSCs vs. E/F MIAMI b. Na[unreadable]ve vs. Resveratrol pre-conditioning in ex vivo organotypic cultures c. Resveratrol pre-conditioned in asphyxia-induced cardiac arrest animal model Aim 2: Comparative effects of cell conveying during the acute (60-min) and sub-acute (24-h) settings a. Injected cells @ 60-min (acute) vs. 24-h (sub-acute) b. Cells conveyed in unloaded laminin-coated PAMs @ 60-min (acute) vs. 24-h (sub-acute) c. Cells conveyed in unloaded laminin-coated PAMs loaded with Resveratrol @ 60-min (acute) vs. 24-h (sub-acute) PUBLIC HEALTH RELEVANCE: Cerebral ischemia affects millions of Americans every year. Current treatments, although helpful, fail to prevent cognitive, motor, and speech impairment due to brain damage caused by cardiac arrest. This proposal focuses on developing cell-based therapies for cerebral vascular diseases offering a new paradigm for advancing and transforming the care of patients suffering global cerebral ischemia. The long-term goal of our labs is to prevent neurological consequences of cardiac arrest. In the current proposal we will focus on preventing hippocampal CA1 neuronal death due to cardiac arrest using a subpopulation of highly homogeneous mesenchymal stromal cells (MSCs) termed marrow-isolated adult multilineage inducible (MIAMI) cells.